To properly address the project objectives and work plan, the
EnzOx2 consortium brings together three
highly-specialized SMEs in the areas of special (basidiomycete)
enzymes (JenaBios), 5-hydroxymethylfurfural production and management of its properties
(AVA Biochem) and chiral chemicals and active pharmaceutical ingredients
(Chiracon); two world-leading companies in the sectors of industrial
enzymes (Novozymes) and flavour & fragrance
ingredients (Firmenich); one technological
centre dedicated to the plastics sector (AIMPLAS);
and six research/academic partners with high expertise in oxidoreductase
structure-function and engineering (CIB
and ICP),
their application in different biotransformations (TUDresden
and IRNAS) and the optimization of enzyme-based bioprocesses
(TUDelft) and LCA analysis and biocatalyst immobilization
(USC).

The latter academic and research partners will contribute to enzyme
discovery and improvement (using protein engineering tools) and will
develop the enzymatic technologies at the lab scale, using the enzymes
provided by Novozymes and JenaBios and those obtained at their own labs.
TUDelft will contribute to the final set-up of reaction conditions and
downstream processing by the enzyme-using companies (AVA Biochem,
Firmenich and Chiracon), who provide the enzyme substrates
to be used by the other partners. Finally, all the companies, together
with the plastic technology centre and the group from USC, will contribute to the evaluation of
the technical (economic and environmental) feasibility of the new bio-chemical
technologies developed, and their integration into lignocellulose biorefineries.

Angel T. Martínez is Research Professor of CSIC
and co-leader of the group of Biotechnology for Lignocellulosic Biomass of
the Environmental Biology Department at the CIB. He is member of ACS, ASM
and TAPPI, with around 300 publications mainly
related to oxidoreductases and microbial biotechnology. He has coordinated
several projects of Industrial Biotechnology from FP3 to FP7, such as
INDOX (KBBE-2013-7-613549),
PEROXICATS (KBBE-2010-4-265397),
BIORENEW (NMP2-CT-2006-026456),
PELAS (QLK3-1999-00590),
PITCH (QLK5-1999-01357)
and AIR2 CT93-1219.

EnzOx2 Partners

Click on the map locations or in logos to access information on the partners

Description of the group
The group of
Biotechnology for Lignocellulosic Biomass
has wide expertise in microbial
enzymes of biotechnological interest. It is part of the Environmental
Biology Department of the CIB, the largest Biology
institute of the Spanish Research Council (CSIC).

Marta Pérez-Boada

Susana Camarero

The contribution to EnzOx2 will be led by Dr
Francisco J. Ruiz-Dueñas, CSIC Tenured Scientist, who
has large experience in heme peroxidases (molecular biology, structure-function
and engineering) resulting in ~65 publications.
Dr Susana Camarero, CSIC Tenured Scientist with
~70 publications devoted to ligninolytic fungi and oxdoreductases
for biotechnology applications will also be involved in the project.
Dr Marta Pérez-Boada, Project Manager with scientific background
in the field of heme-peroxidases, will be in charge of the
Project Secretariat (see Contact section) assisting in project
coordination, management and dissemination.

Main role in EnzOx2
The main contributions of CIB will be in oxidoreductase
engineering and in the development of an enzymatic technology for the
production of sugar-derived building blocks. Minor contributions are
expected also for some specific enzymes and techniques available at CIB.

Angel T. Martínez

Project Coordination will be performed at CIB by Prof
Angel T. Martínez. The Coordinator and Project Secretariat will be
assisted by the CSIC European Programme Unit in those
aspects related to Consortium Agreement management and project administrative coordination.

Description of the group
The group belongs to the Department of Plant Biotechnology at IRNAS
(one of the CSIC institutes in the area of Agriculture),
and has two senior members, Dr Ana Gutiérrez and Dr
José C. del Río who have high expertise in the chemical
and structural characterization of lipids and lignin by different analytical
techniques. Additionally, they have expertise in the evaluation of the structural
modifications of these organic fractions after industrial processing, and
chemical or enzymatic treatments.

The IRNAS contribution to EnzOx2 will be led by Dr
Ana Gutiérrez, CSIC Research Scientist, current Vice-director
of IRNAS, recent elected member (2015) of the International Academy of Wood
Science (IAWS) and of the IAWS Academy Board (2016), who has ~130 SCI articles
and has participated in the coordination of 5 EU projects. Other members of the
group are Dr Jorge Rencoret (with expertise in 2D NMR,
thioacidolysis and GC-MS) and several PhD students.

Main role in EnzOx2
The main role will be the study of the reactions of oxyfunctionalization
(hydroxylation) of aliphatic compounds (basically saturated/unsaturated
fatty acids, and terpenes/terpenoids) using peroxygenases. The products
will be analyzed mainly by GC and GC-MS using short- and medium-length
high-temperature capillary columns. Additionally, pre-treatment reactions
using laccases will be investigated by following the changes produced in
the lignin polymer using 2D NMR, among other techniques. IRNAS
will also contribute to management in WP7 contributing
to Dissemination of results.

Description of the group
ICP is the main institute of the catalysis area of the Spanish CSIC.
The Directed Enzyme Evolution group of the CSIC Institute of Catalysis is
led by Miguel Alcalde, and primarily focuses its research
on the engineering of enzymes for a wide range of biotechnological purposes;
the development of high-throughput biomolecular screening tools and genetic
methods for library construction and exploration, as well as synthetic biology
studies for environmental and energy applications. The laboratory has participated
in several consecutive European programs and research contracts with industries
with several patents and ~70-SCI papers in the field.

The ICP Team.

Main role in EnzOx2
The ICP contribution will be dedicated to improve some of the selected enzymes
(AAO and peroxygenase) for specific target reactions (oxidation of FFCA,
and terminal hydroxylation of fatty acids, respectively) using molecular
directed evolution approaches.

Description of the group
The International Institute Zittau (IHIZ), a former small state university
in Saxony in the triangle where Germany, Poland and the Czech Republic meet,
has been, since January 2013, a Central Academic Unit of the Dresden University
of Technology (TUDresden), the only excellence university in East Germany.
IHIZ was founded in 1993 and is especially dedicated to the joint education of
German and foreign Master and PhD students (International Institute Zittau).

Mycologists of the TUDresden during field campaign – R. Ullrich and A. Karich
searching for rare, lignocellulytic fungi as new enzyme producer on dead
wood in guidance with Claus Bässler in the Bavarian Forest National Park
(Germany, Photo: H. Kellner)

The Department of Bio- and Environmental Sciences consists of five research
groups (Mycology & Enzymology, Bioinorganic Chemistry, Environmental Chemistry,
Environmental Process Engineering, Hydrology) and has, at the moment, 26 members
(3 technicians, 4 Master-, 13 PhD students, 2 postdocs, 2 senior scientists,
2 professors). Moreover, more than 50 visiting scientists from 14 countries
have worked over the last years at the department.

Ceratiomyxa fruticulosa (Photo: A. Karich)

The research line of the department deals with the functional biodiversity
of microbial enzymes, with special emphasis on the biodegradation of natural polymers
and the oxyfunctionalization of organic molecules. During the last years,
the scientific interest has focused onto peroxygenases, their catalytic
cycle, molecular background and eco-physiological relevance.

Main role in EnzOx2
TUDresden will focus on fungal peroxygenases, their selection, performance in the conversion
of HMF to FDCA and in the oxyfunctionalization of terpenoids including steroids as well as
on molecular aspects of their stability. Furthermore TUDresden will collaborate with JenaBios
in the production of wild-type peroxygenases and AAOs.

Description of the groupJenaBios distributes fungal peroxidases and phenoloxidases
for biotechnological applications, research and development.
It was founded in 1998 by scientists who wanted to share their proven utilities
and experience with the life science community. The company is located in the
“High-tech valley” of the traditional university town Jena in Germany and is
integrated in a consortium of 3 companies (with more than 20 academics)
with several business areas and product fields. Enzymes are produced by
fermentation of highly selective fungi and purchased as bulk or partly purified
biocatalysts. Applications of ligninolytic enzymes and fungi are oxidative
syntheses, degradability tests, modification of natural and synthetic materials,
waste treatment in the textile and pulp-paper industry, coloring processes,
bioremediation (e.g. of organo-arsenical warfare agents and explosives),
oxidative degradation and bleaching processes and nutritional modification
of animal feed.

Main role in EnzOx2JenaBios will extend the product portfolio and provide oxidoreductases,
e.g. peroxygenases and oxidases, in several purities and immobilization stages.
Together with TUDresden, K. Scheibner first described peroxygenases
and will develop during EnzOx2 very specific production and
isolation strategies. Several biotechnological application perspectives
of peroxygenases were patented/described by JenaBios.

Description of the groupFirmenich is the largest privately-owned company in the perfume
and flavor business. Swiss and family owned, we have created
many of the world’s favorite perfumes for over 100 years and produced a number
of the most well known flavors we enjoy each day. Our consistently superior
investments in R&D substantiate our desire to understand, share and reinvent the
best of what nature has to offer in the realms of smell and taste.
Firmenich has significant expertise in the areas of biocatalysis,
synthetic biology and terpene biochemistry. Furthermore, the company has a
strong track-record in the development of biochemical transformations to access
both perfumery and flavor ingredients. Dr. Andreas Taglieber,
who is in charge of a research team developing biotechnological technologies
and processes to access flavour and fragrance ingredients in Firmenich, will coordinate
the resources and personnel involved in EnzOx2.

Main role in EnzOx2Firmenich will contribute to the project by proposing and supplying
substrates that will permit to develop enzymatic transformations which are
valuable to the flavour and fragrance industry. It will also perform screenings and
further testing of promising enzymes that will be discovered by other project
partners and supplied to Firmenich, followed by technical/economical
evaluation of the best candidates.

Description of the groupAVA Biochem, a SME founded in 2012, is a subsidiary of AVA-CO2 Switzerland,
founded in 2009. AVA Biochem developed the conversion from biomass
to 5-HMF to industry scale. It managed to set up a reliable process and produce
a consistent quality of 5-HMF. AVA Biochem managed to stabilize 5-HMF, which
was a major problem in the past. A major breakthrough was reached in terms
of economic feasibility: the process design makes large scale production
(30,000 – 100,000 t/a) possible at a price which is close to comparable
petro-based product. Right now, AVA Biochem is selling crystallized 5-HMF
in purities of 95-99% on a small scale to research institutions worldwide.

Gilbert Anderer will be the person responsible for the AVA Biochem
contribution. Other members of the company involved in EnzOx2
will be S. Krawielitzki, V. Wagner-Solbach, J-U Wichmann, F. Badoux and M.-J. Desroches.

François Badoux optimally combines his economic talent with his
knowledge in chemical engineering. He began his career in 1983 at Ciba-Geigy and
was then a consultant for 4 years with McKinsey & Co., where he specialized in
industrial processes and strategies. Then, he led the International Engineering
Consultancy Department at Sete SA, a large private industrial and finance group.
After 12 years, he founded Mistral Engines S.A., an aerospace company. There he
developed a new generation of light aircraft and helicopter engines to market readiness.
François Badoux has a degree in chemistry from the ETH Lausanne and an MBA from the IMD.

Main role in EnzOx2
The main contributions of AVA Biochem will be produced in the supply
of different HMF qualities and evaluation of biomass separation/pretreatment technologies,
the comparison of these HMF preparations in the enzymatic production of FDCA,
and the technical and economic evaluation of the enzymatic technology for FDCA
production from HMF, compared with other relevant chemical and microbial technologies.

NovozymesNovozymes is world leader in bio-innovation and the world largest
producer of industrial enzymes, with a 48% market share. Its over 700
products are used in more than 40 industries to improve processes and quality,
and save water, energy, raw materials and waste. In 2015 Novozymes reported
sales of 1881 million € and spend around 14% of its turnover on R&D.
It is headquartered in Denmark and employs more than 6000 people in 30 countries,
with production plants in Denmark, US, China, Brazil and India.

Description of the group
The Technical Industries R&D group is responsible for enzyme product and process
development within a range of industries, including pulp & paper, waste water,
leather and biogas. The group (headed by Erik Gormsen) has around
25 people dedicated to the above areas with broad expertise in the field of industrial
enzymes. Owik Herold-Majumdar has worked with oxidoreductases,
in particular peroxygenases, on designing novel bioanalytical synthesis routes for
bulk and specialty chemicals. He will be the key industrial contact and coordinating
the enzyme sampling along with assisting in enzyme reaction design.

Main role in EnzOx2
The Novozymes contribution will consist in production of different
wild-type and engineered enzymes (such as fungal peroxygenases, oxidases, laccases,
cellulases/xylanases and isomerase) and their supply to research/academic partners
for development and optimization of enzymatic technologies and other companies for
their evaluation in the manufacture of renewable polyesters, and some specific F&F
ingredients and APIs, among other added value compounds.

Description of the group
The ‘Biocatalysis Group’ of TUDelft headed by
Prof. W.R. Hagen is dedicated to Biocatalysis and Organic
Chemistry aims at discovery, design and engineering novel enzymatic pathways
leading to target products. To strengthen the focus in the application-oriented
research, we have formulated the understanding of the effect of immobilization
(stability as well as compartmentalization) of enzymes as a central research line.
Themes and topics for specific research projects are inspired by our contacts
with industrial partners, as well as contacts with academic colleagues in Europe
and world-wide.

Dr. Frank Hollmann is an internationally renowned expert for
the use of enzymes in organic synthesis. His research interests
span diverse classes of oxidoreductases for preparative oxidation/reduction
and oxyfunctionalization chemistry. The infrastructure of BOC
can be described as a combination of base organic chemistry and molecular
biology laboratories. In addition, spectroscopy facilities, such
as NMR, and EPR are present within the group. For screening of enzymes,
a dedicated laboratory is available, that allows performing high-throughput
production and assaying of enzymes at advanced safety levels (ML2).

Main role in EnzOx2Dr. Hollmann will be the scientist in charge (supported
by staff of the Department of Biotechnology). The group will be
involved in the optimization of the enzymatic reactions towards industrial
application. They will act as facilitator between the groups
discovering and optimizing new enzymes and the industrial partners wanting
to commercialize them. Both, Prof. Hagen and Dr. Hollmann
will be actively involved in the project, together with Dr. Florian Tieves,
project contract at the BOC group.

Description of the groupChiracon is an experienced and independent, privately
owned company with in-house technology comprising of pharmaceutical
and analytical laboratories. The CEO of the company is Dr. Ralf Zuhse.
The company was founded 1998 in Luckenwalde, close to Berlin.
The activities of Chiracon are focused on RD, up-scaling
and manufacturing of advanced chiral intermediates and active pharmaceutical (APIs)
under Good Manufacturing Practice (GMP) conditions for the pharmaceutical
industry. The team has four senior scientists, Dr. Thomas von
Schrader (chemistry, scale-up), Dr. Stephan Quint
(chemistry, development), Dr. Simon Michaelis
(chemistry, development) and Dr. Ahmad Tahrani (Pharmacy,
analytics and quality control). Chiracon is certified
according ISO 9001 since 2000. Serving as a bridge between medicinal
chemistry and full-scale manufacturing, Chiracon provides
process research services with a core competency in the rapid supply of
APIs according to GMP standards. Due to almost 20 years of solid experience in the synthesis
of chiral and non chiral APIs for early phase drug development, Chiracon can support
their customers through pre-clinical and clinical trials.

Main role in EnzOx2Chiracon will supply steroid raw materials and
products/intermediate standards, and provide analytical support,
contribute to reaction and downstream optimization, and perform the
technical and economic feasibility evaluation for the enzymatic
production of a pharmaceutically active ingredient.

Description of the group
The Group of Environmental Engineering and Bioprocesses
belongs to the Department of
Chemical Engineering of USC. It can be considered one of the best Environmental Engineering groups in Spain,
focusing on four main topics: (i) Environmental Management: Clean technologies and Life Cycle Assessment;
(ii) Development, monitoring and control of technologies for wastewater treatment, (iii) Technology transfer
for wastewater treatment and bioprocesses, (iv) Ligninolytic enzymes for degradation of recalcitrant compounds.

The following people will be involved in the EnzOx2 project: Mª Teresa Moreira (professor with >
210 JCR publications, h-index of 41 and Associated Editor of Journal Cleaner Production); Gumersindo Feijoo
(professor with >200 JCR publications and h-index of 42), Yolanda Moldes (PhD student) and
María Gamallo (PhD student).

Main role in EnzOx2USC will focus on the immobilization of oxidative enzymes (UPOs, laccase, versatile peroxidases)
in different supports and will perform the sustainability assessment of the different production processes
developed in the framework of the project.

AIMPLAS, a Plastic Technology Center located in Valencia,
is a private, non-profit Association with about 450 associated companies
(the majority of them SMEs). AIMPLAS has offered its services to the Plastic
industry sector since 1990 and, nowadays, has a permanent staff of 110 people.
AIMPLAS’ fields of work are related to technological research and development
on plastic materials & products and its transformation processes.

Description of the group
The team has mutidisciplinar senior scientists formed by Dr. Amador
García-Sancho, PhD in Organic Chemistry and BD in Pharmacy by the
University of Valencia, Dra. Alba M. Ortiz, European PhD
in Chemistry by the Universidad de Alcalá, Dra. Belén Monje,
Ph.D. in Organic Chemistry by the University of Valencia, Ms. Itziar
Carracedo, Chief Project Manager and Ms. Marta Estellés,
Projects Manager. All of them have a wide experience in the synthesis
and characterization of polymers and new polymers taking always into account
the appropriate cost effectiveness of the polymer synthesis process, optimizing
parameters as reaction time, temperature profile, amount of catalyst as well
as the post-purification steps.

Main role in EnzOx2
The main contribution of AIMPLAS will be in the techno-economic
evaluation of the enzymatic technologies developed by other partners in: i) the
synthesis of chemical building blocks (such as FDCA and hydroxylated fatty acids);
and ii) in the subsequent production of different polyesters (bioplastics)
from these building blocks.